1-(2-alkyl-4-amino-5-pyrimidinylmethyl)-alkyl-pyridinium quaternary salts for treating coccidiosis



This invention relates generally to new chemical compounds. More specifically, it is concerned with chem; ical compounds which are useful in the prevention and the cure of coccidiosis Still more particularly, it is concerned with a series of new pyridinium quaternary salts which are effective in controlling coccicliosis when fed in small amounts to poultry; Still more specifically. it relates to 1- (2-loweralkyl-4-amino S- yrimidylmethyl)- loweralltyl-pyridiniurn quaternary salts where the lower alkyl radical in the pyrimidine ring contains at least twov carbons, and with methods for preparing such compounds. It is further concerned with novel con'rpositions containing such substances.

This application is a division of our copendingap plication Serial No. 829,979 filed July 28, 1959, which 1- in turn is a continuation-in-part of our application Serial No. 805,688, filed April 13, 1959; now abandoned.

Coccidiosis is a common and widespread poultry disease caused by several species of protozoan parasites of the genus Eimeria, such as E. tenella, E. necatrix, E. acervulina, E. maxima, E. hagzmi and E, brunerti. E renella is the causative'agent of a severe and often fatal infection of the ceca of chickens which is manifested by extensive hemorrhage, accumulation of blood in the ceca, and the passage of blood in the droppings E. necatrix as well as certain other species attack the small intestine of the chick causing what is known as intestinal coccidiosis. Related species of coccidia such as E. malagrizlis and E. adenoides are causative organisms of coccidiosis in turkeys. When left untreated, the severe forms of coccidiosis lead to poor weight gain, reduced feed efficiency and high mortality in fowl. The elimination or control of coccidiosis is, therefore, of paramount impor tance-in the poultry raising industry.

It has now been found that certain pyridiniuin quaternary compounds are very highly active against the protozoa responsible for coccidiosis. An object of the present invention is to provide such novel compounds. Another object is to provide a synthesis 'of such substances. A further object is the provision of animal feeds and feed supplements and of water soluble compositions containing these pyridinium quaternary salts. Other objects will be apparent from the following discussion of our invention.

According to this invention it has been found that 1- (2-loweralkyl-4-amino 5 pyrimdylmethyl) loweralkylpyridiniurn quaternary salts, wherein th'e pyridine ring is substituted with from one to three lower alkyl groups and the lower alkyl radical at the 2-position of the p' y' rirnidine ring contains at least two carbon atoms, are very effective in preventing and treating coccidiosis. The preferred compounds of our invention are quaternary salts of the formula wherein R is a lower alkyl radical containing at least tes atent O 7 3,22% assented Feb. a, less ice two carbon atoms, R is a lower alkyl radical, n has a value of 1, 2 or 3, and X is an anion; b and c are positive numbers having values such that the positive charge of 17 moles of cation is neutralized by c moles of anion X. Thus when X is a mono'v ale nt anion such as a halide, b is l and c is 2. When more than one lower alkyl group is present in the pyridine ring, i.e. when n is 2 or 3 such lower alkyl groups may be the sameor different. As described more fully below, these compounds are prepared by reaction of a 2-loweralkyl-4-amino-S-substitutedmethyl pyrimidine with an appropriately alkylated pyridine. As will be apparent from the above structural formula, the compounds described herein may be considered substituted pyridines. The pyridine ring is substituted at the 1position by a 2 -loweralkyl-4-amino-5- pyrimidylmethyl radical; it is further substituted with one or more lower alkyl groups such as methy l,ethyl prqpyl, is'o rbpy l, butyl amyl groups, such substituents being in at least one of the 2, 3 and/ or 4 positions of the ring. The pyrimidine moiety also contains a lower alkyl group at the 2'position of the pyrimidine ring, which groupis one containing at least two carbon atoms, e.g. ethyl, propyl, isopropyl, isobutyl and the like. The lower alkyl group present in the pyrimidine and pyridine portions of these salts need not, of course, be the same in any particular compound. 7 I p 7 Although the compounds of the invention are active generally against coccidiosis, certain of the quaternary salts are particularly advantageous in that they possess a broader.anticoccidial spectrum, i.e., are active at low levels against more of the most troublesome strains of coccidia, than others, and in that they exhibit minimal undesirable side eifects. The preferred compounds are the 1-(2-ethyl or Z-propyl-4-amino-S-pyrimidylmethyl)- loweralkyl pyridinium quaternary salts having a lower alkyl radical at either the 2 or 4 position of the pyridine ring. Included among these are the l-(2-ethyl-4-a'rnino- 5-pyrirnidylrnethyl) -2-methyl pyridinium salts, 1-(2-ethyl- 4-arnino-5-pyrimidylmethyl)-4--methyl pyridinium salts, 1 (2 propyl-4-afnino-5-pyrimidylrnethyl)-2-methyl pyridiniurn salts, and l 4 (2 propyl-4-amino 5 pyrimidyl rhethyl)-4-methyl pyridiniurn salts, which substances e hibit a very favorable spectrum of anticoccidial activit' and therapeutic index. 1-(2-butyl-4-arnino-5-pyrimidylmethyl)-3-methyl pyridiuium salts are likewise preferred since they are particularly eifective against E. ncervtllin'a.

The anion of the quaternary compound may be an in organic anion such as chloride, bromide, iodide, nitrate, sulfate, phosphate and th'elike, or the anion of an organic acid such a's'citric, tartaric, acetic, picric, stearic, su'ccini'c, benzoic, phthalic, phenoxyacetic, emboni'c, abieti'c'; Z-naphthalene sulfonic or ethylenediamine tetracetic acids. It may also be the anion of a polymer such as a polyphosphateor p'olys'tyrenes'ulfonate ion. The nature of the anion is-not critical and any anion may be employed as long as it is not unduly toxic for poultry when the compounds of the invention are employed as coccidiostats. It will be realized by those skilleld in this art that an" acid addition salt of the primary amino group present in these conipouridswill also be formed concurrently with the quaternary salt. I, I

The compounds of this invention are synthesized by reacting together an appropriately substituted pyrimidine and a lower alkylated pyridine. As the pyrimidine reactant we may employ an ester of a 5-hydroxymethyl pyrimi dine and a strong inorganic a 'd such as a hydrohalic acid; According to one process, a 2-loweralkyl-4 amino- S-halon'aethyI pyrimidine dihydrohalide, in which the halogen is bromine or chlorine and the loweralkyl radical contains at least two carbon atoms, is reacted directly with the alkylated pyridine. An excess of the liquid alkylated' pyridine or, alternatively, organic solvents inert under the CH3X where R is lower alkyl having at least two carbons and R is a lower alkyl radical, n is 1-3 inclusive and X is a halogen such as chlorine or bromine.

Although the S-halomethyl pyrimidines are generally most conveniently employed for reaction with the alkyl pyridine, the quaternization may also be brought about with other esters of the 2-loweralkyl-4-amino-5-hydroxymethyl pyrimidine such as the nitrate ester. Other suitable esters are those of organic sulfinic and sulfonic acids such as the methylsulfinate, p-toluenesulfonate and benzenesulfonate esters.

The quaternization may be conducted so that the particular salt desired is obtained directly. Mternatively, the quaternary salt recovered from the synthetic reaction medium may be conveniently metathesized to another salt by techniques known in the art.

In using the compounds of this invention in the treatment and prevention of coccidiosis, they are conveniently fed to poultry as a component of the feed of the animals although they may also be given dissolved or suspended in the drinking water. According to one aspect of the invention, novel compositions are provided in which compounds of Formula I above are present as an active anticoccidial ingredient. Such compositions comprise the quaternary salts intimately dispersed in or admixed with an inert carrier or diluent. By an inert carrier is meant one that is nonreactive wtih respect to the quaternary and that may be administered with safety to the animals. The carrier or diluent is preferably one that is or may be an ingredient of the animal feed.

The compositions which are a preferred feature of this aspect of the invention are the so-called feed supplements in which the active ingredient is present in relatively large amounts and which are suitable for addition to the poultry feed either directly or after an intermediate dilution or blending step. Examples of carriers or diluents suitable for such compositions are solid orally ingestable carriers such as distillers dried grains, corn meal, citrus meal, fermentation residues, ground oyster shells, Attapulgus clay, wheat shorts, molasses solubles, corn cob meal, edible vegetable substances, toasted dehulled soya flour, soybean mill feed, antibiotic mycelia, soya grits, crushed limestone and the like. The quaternary salts are intimately dispersed or admixed throughout the solid inert carrier by methods such as grinding, stirring, milling or tumbling. By selecting proper diluents and by altering the ratio of carrier to active ingredient, compositions of any desired concentration may be prepared. Formulations containing from about 1% to about 40% by weight, and preferably from about 2-25% by weight of active ingredient are particularly suitable for addition to poultry feeds, and compositions containing from about 5-15 by weight of coccidiostat are very satisfactory. The active compound is normally dispersed or mixed uniformly in the diluent but in some instances may besorbed on the carrier. The optimal concentration of coccidiostat in these feed supplements will depend to some extent on the particular compound employed. Since it is convenient for the feed manufacturer to use about one pound of feed supplement for each ton of finished feed, the preferred concentration of any one of our coccidiostats in a feed supplement is partly a function of the level of active ingredient desired in the finished feed.

Examples of typical feed supplements containing a 1-(2-loweralkyl-4-amino 5 pyrimidylmethyl) loweralkylated pyridinium quaternary salt (where the lower alkyl radical in the pyrimidine ring has at least two carbon atoms) dispersed in a solid inert carrier are:

Lbs. l-(2-ethyl-4-amino-5 pyrlm' idylmethyl) 2 methyl pyridinium chloride hydrochloride 6.0 Wheat standard middlings 94.0

1-(2-ethyl-4-amino 5 pyrimidylmethyl) 4 ethyl pyridinium bromide hydrobromide 10.0 Corn distillers dried grains 90.0

l-(2-ethyl-4-amino-5-pyrimidylmethyl) 2 methyl- S-ethyl pyridinium chloride hydrochloride 15.0 Wheat standard middlings 85.0

1-(2-ethyl-4-amino 5 pyrimidylmethyl) 2,3 dimethylpyridinium bromide hydrobromide 12.0 Molasses solubles 88.0

1- 2-n-propyl-4-amino-5-pyrimidylmethyl) -2 -methyl pyridinium chloride hydrochloride 20.0 Corn germ meal 30.0 Corn distillers grains 50.0

F. 1-(2-ethy1-4-amino-5 pyrimidylmethyl) 2 methyl pyridinium embonate 40.0 Soya grits 60.0

1-(2-ethyl-4-amino-5-pyrimidylmethyl)-4-n propylpyridinium sulfate 7 Fermentation residues 50.0 Wheat shorts 45.0

H. 1- 2-n-propyl-4-amino-S-pyrimidylmethyl) -4 -methyl pyridinium bromide hydrobromide 12.0 Wheat standard middlings 88.0

These and similar feed supplements are prepared by uniformly mixing the quaternary salt with the carrier or carriers.

The feed supplements of the type illustrated hereinabove are usually further diluted with materials such as corn meal or soybean meal before beingincorporated in the animal feed. In this intermediate processing step the level of coccidiostat in the carrier is brought down to from about 0.1% to about 1.0% by Weight. This dilution serves to facilitate uniform distribution of the substance in the finished feed. The finished feed is one that contains a source of fat, protein, carbohydrate, minerals, vitamins and other nutritional factors.

The amount of pyridinium quaternary salt required for optimum control of coccidiosis in poultry will, of course, vary somewhat with the specific compound or compounds employed. The compounds of Formula I above are effective when administered at levels of less than about 0.05% by Weight of the feed. With the preferred compounds of the. invention, i.e. the 1-(2 ethyl-4-amino 5 pyrimidyh' methyl)-2(or 4)'-loweralkyl pyridiniurn and 1-(2-propyl- 4-amino-S-pyrimidylmcthyl)-2(or 4)-loweralkyl pyridinium salts, good prophylactic results are obtained when from about 0.0005 to about 0.05% by weight of the total feed consumed is administered; for most satisfactory results from the standpoint of both efiicacy and incidence of undesirable side effects it is preferred that the poultry iced contain between about 0.0025% and 0.025% by weight of pyridiniurn salt. When the pyridinium salts are to be employed as therapeutic agents, the higher levels may be used for relatively short periods of time. Thus, concentrations of about 0.02% to 0.05% by weight of the feed may be advantageously administered in treating an established outbreak of coccidiosis. It is desirable to employ the lowest levels that afford fully adequate control of coccidiosis in order to eliminate as far as possible any risk of side eifects that might appear on prolonged feeding of the compounds.

Many of these pyridinium quaternary salts are desirably or advantageously administered to poultry by way of the drinking water of the birds. This method of treatment is often employed in the therapeutic use of our cornpounds since poultry with coccidiosis are apt to consume less solid feed than normal birds. The water=soluble quaternary salts may be added directly to the drinking water. Alternatively, water-soluble powders may be prepared, in which the coccidiostat is intimately admixed with a suitable carrier, such as dextrose or sucrose, and these powders added to the drinking water of poultry as necessary. Such water-soluble powders may contain any desired concentration of coccidiostat, and preparations containing from 125% by weight of active compound are suitable.

The following examples are given for purposes or illustration and not by way of limitation:

EXAMPLE 1 A. A slurry of 100 grams of 2-ethyl-4-amino-5-bro1nomethylpyrimidine dihydrobromide in 500 ml. of acetonitrile was stirred at room temperature and 100 m1. of 2-methyl pyridine (Ot-PlCOllHB) was added as rapidly as possible. The solution became almost clear, and the temperature rose from 25 C. to 35 C. 1-(2-ethyl-4- amino-S-pyrimidylmethyl)-2-methyl pyridinium bromide hydrobromide started to precipitate in about a minute. The reaction mixture, after stirring overnight at room temperature under nitrogen, was filtered, the solid Washed with 100 ml. of acetonitrile and two 100 ml. portions of ether, and dried under vacuum at 40 C. The 1-(2-ethyl-' 4-amino-S-pyrimidylmethyl) 2 methyl pyridinium bro mide hydrobromide thus obtained weighed 99 grams, melting point 266270 C. (dec.).

B. A slurry of 600 ml. of Amberlite Hi -45 resin in two liters of 2.5 N hydrochloric acid was put in a column and backwashed with distilled water to remove fine solids. The resin was then washed with distilled water until the eluate was less than 0.05 N in hydrochloric acid. The pyridinium bromide obtained in part A. above was dissolved in two volumes of water and the solution put on the resin column. The flow rate was adjusted to 15 ml.

per minute, and after a displacement cut of about 300 ml.,

three product cuts of 200 ml. each were taken. The presence of product in the eluate was indicated by color, striations, and halide titer; The three product cuts were combined when potentiometric titration with silver nitrate indicated the absence of bromide ion, and concentrated under vacuum to a slurry of about ml. The slurry was stirred at room temperature and diluted with two liters of acetone. After aging for two hours in an ice bath, the 1-(2-cthyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride was filtered ofi, washed with two 100 ml. portions of acetone and dried under vacuum at 40 C. Yield 73 grams, 95% of theory. U.V. (0.1 M HCl) r 2460, E% 448; A 26%, E% 433. Melting point zen-267 C. (dec.).

6 EXAMPLE 2 To '5 of :Z-methylpyridin'e (a-picolin'e) was added a solution of -2 grams of -2-ethyl-4 amino'-5-pyiiinidylmethyl bromide dihydrobromide in 15 ml. of methanol. A clear solution formed which became slightly warm. in a short time crystals of 1-(2 ethyl-4 amino 5-pyrimidylmethyl)-2-methylpyridinium bromide hydrobromide began to appear. The mixture was allowed to stand for about 15 hours and then a mixture of ether-petroleumether added to complete the crystallization. The crystalline material was then collected by filtration, and re'crystal lized from 18-20 ml. of methanol. The crystals of l-(2- ethyl-4 amino-S-pyrimidylmethyl)-2 methylpyridinium bromide hydrobi'omide thus obtained melt at about 270 C. (dec.).

EXAMPLE 3 One gram of 2-propyl-4-amino' 5-pyrirnidylmethyl bromide dihydrobromide was dissolved 'or suspended in 7 rnl of drly dimethylforinamide, and to this solution was added 5 ml. of Z-methyl pyridine. A clear solution formed from which crystals of 1-(2 ropyi -4 amino S-pyrimidyI- meth'ylIt-Z-rriethyl pyridiniui'n bromide hydrobromide soon precipitated. These were collected by filtration and dried, melting point 242-3 C. (dec.). The material was purified by recrystallization from a methanol-ethanol mixture. EXAMPLE 4 To a suspension of 7.6 gra n's of Z-ethyl-4-amino 5- pyrimidylmethyl bromide dihyd'robromide in 50 ml. of acetonitrile was added 9 ml. 'of 2 m' ethyl-5ethyl pyridine. This mixture became warm and a clear solution resulted. After a short time 1-(2-ethyl-4amino-Spyriruidylmethyl) 2-mcthyl-5-ethyl pyridinium bromide hydrobromide beg'an to crystallize. Crystallization was continued overnight and the white crystals then collected by filtration, melting point 252 C. (dec.). The crystals were dissolved in 5 0 ml. of methanol and the solution then diluted with ml. of acetone. The crystals of product thus obtained melted at 261 C. (dec.).

EXAMPLE 5 1.9 grams of p-toluene sulfon'yl chloride was added gradually with shaking to a cooled (0-5" C.) solution of 1.67 grams of 2-n-propyl-4-a'mino-5-hydroxy-methyl pyrimidine in 10 ml. of 2-methylpyridine. The reaction mixture, after standing three hours in an ice bath, and 15 hours at room temperature, was evaporated to dryness in vacuo. The residue was dissolved in 20 ml. of water,- acidified with hydrochloric acid and poured over a column of Amberlite IRA-400 ion exchange resin on the chloride cycle. The eluate was evaporated to dryness to give a residue of 1-(2-n-propyl-4-amino-5pyrimidylmethyl)-2- methyl pyridinium chloride hydrochloride. On recrystal lization from a methanol-ethanol mixture, the quaternary had melting point 246 C. (dec.).

EXAMPLE 6 200 mg. of 1-(2 ethyl-4-amino S-pyrimidylmethyl)-2- methyl pyridinium bromide hydrobromide was dissolved in 0.8 ml. of concentrated hydrochloric acid. This solution was carefully diluted with acetone until 1-(2-ethyl-4- amino-5-pyrimidylrnethyl)'-2-methyl pyridiuium chloride hydrochloride crystallized. This salt was dissolved in 0.5 ml. of concentrated hydrochloric acid and the solution slowly diluted with about 8 ml. of acetone. The salt crystallized and on drying melted at 279-281 C. (dec.).

EXAMPLE 7 5.1 grams of 2-ethyl-4-amino=5-chlorornethyl pyiirni dine hydrochloride and 25 ml. of 2-met hyl pyridine were mixed together in a 50 ml. flask and the mixture warmed on a steam bath, with stirring for two hours. The mixture was then allowed to stand at room tempera ture for about 15 hours. The solid 1-(2-ethyl-4-arnino-5- pyrimidylmethyl) -Z-methyI pyridiniurn chloride hydfo- EXAMPLE 8 compounds of the invention was determined by the following method:

Groups of ten two-week old white leghorn chicks were fed a mash diet containing various amounts of the com- 5 pounds. The compounds were uniformly dispersed in the The pyridinium quaternary salts set forth in Table I feed. After having been on this ration for 24 hours, each below were synthesized utilizing processes described in chick was inoculated with 50,000 sporulated oocysts of preceding examples. In most experiments a 2-loweralkyl- E. lenella. 4-amino-5-halomethyl pyrimidine was employed as the Other groups of ten chicks each were fed a slrmlar pyrimidine reactant, although in some cases an ester of 10 mash diet which contained no coccidiostat. These were a S-hydroxymethyl pyrimidine was utilized. also infected after 24 hours and served as positrve or Tablel NHz.2HX R R N1=n+ R, R

N orms, .214121,- -nr 22:-

i I R,KN/ Ra Rs k R R3 Quatern- Cpd. ary pre- Melting Percent No. R1 R2 R R R5 R X pared by point 0 in feed method of (100.)

Ex. No.

Br. 2 200-270 0. 001 C1 0 279-281 0.001 Br--- 4 22s 0. 003 C1 0 253 0. 00s C2H5 B1. 4 261 0. 002 C2115 O 0 209 0.002 Br--- 4 244 0.002 Br 4 220 0.001 Br-.. 4 207 0. 000 EL. 4 241 0. 003 CH3 Br Br 4 239 0. 003 Br-- 4 207 0. 000 Bl... 4 252 0. 003 Br- 12 281-288 0. 003 Bra 242-243 0. 003 01 5 240 0. 003 Br-.- 3 244 0.000 0H Ol 0 230 0.01

Br- 12 258260 0. 003 Br- 12 250-200 0. 0125 Br- 12 244-240 0. 000 Br 4 233 0.0125 Br.-- 11 205 0. 0125 Polystyrene sulfonate 0. 004 Embonate 10 0. 003 Naphthalene disulf0nate 10 0. 004

Nora--03 through R are hydrogen unless specified otherwise.

Anticoccidial activity of the pyridinium quaternary infected controls.

salts described in the above table was determined by the following test:

Straight run white leghorn chicks, in groups of three each, were weighed and placed in cages with wire floors. They were fed ad libitum a standard laboratory basal ration in which graded levels of the pyridinium quaternary salt were intimately dispersed or blended. Normal and infected control birds were fed basal ration containing no quaternary salt. On the second day of test, the chicks were each orally inoculated with 50,000 sporulated oocysts of Eimez'ra tenella. Papers under the cages were examined on the sixth, seventh and eighth days of assay for the presence or absence of bloody droppings. A score of 0 was given if no bloody spots were observed; scores of 1, 2 or 4 were assigned for 1-3, 4-6 and 6 bloody spots, respectively. On the eighth day of assay, the surviving birds were weighed, sacrificed and examined grossly for cecal coccidiosis lesions. Normal ceca were scored 0, and ceca with detectable, moderate or maximal lesions were scored 1, 2 and ,4, respectively. When a bird died and cecal coccidiosis lesions were presents, a score of 5 was recorded. If the total of the two scores was 0-5, the compound under test was rated active; if the total score was 6 or more the compound was rated inactive at the concentration tested.

The compounds described in Table I, when tested by this method were found to be highly active (total score of 0-5) at the dose level indicated.

EXAMPLE 9 The coccidiostatic activity of two of the pyridinium As positive controls, two to four groups of ten chicks each were employed. Still other groups of ten chicks each were fed the mash free of coccidiostat and were not infected with coccidiosis. These served as normal controls.

The diets were administered to the chicks for eight days following the date of infection. At the end of this time the infected birds still living were sacrificed. The oocyst count was determined by a microscopic examination of the cecal homogenates.

The results employing the indicated amounts of quaternary salt, and expressed as mean values, are set forth in Table II.

Table II Number Percent Percent Percent of oocysts Compound compound morweight XlO- in in feed tality gain surviving animals 1 (2 ethyl -4-amino -5 -pyrlm- 0. 0015 0 3 ldylmethyl) 2 methyl py- 0.003 0 99 0.1 rldinium chloride hydro- 0. 008 0 106 0.1 chloride. 0. 0125 0 100 0. 1 1 (2 ethyl-4-amino-5-pyrim- 0.0015 0 97 0.2 idylmethyl) 2 methyl 5- 0.003 0 97 0.1 ethyl pyrldiminm chloride 0.006 0 0. 1 hydrochloride. 0. 0125 0 89 0. 1 Infected controls 40 63 6 Normal controls 0 90 EXAMPLE 10 A. Three grams of l-(2-ethyl-4-amino-5-pyrimidy1- methyl)-2-methyl pyridinium bromide hydrobromidein 20 ml. of water was treated with N calcium picrate solution until precipitation appeared to be complete. The solid 1 (2 ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium dipicrate thus obtained was recovered by filtration. It was purified by dissolving in 15 ml. of hot dimethylformamide, filtering said solution, and reprecipitating the salt by treatment of the filtrate with 75 ml. of methanol. n drying the product had a melting point of 1734 C. (dec.).

B. 1 (2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium embonate was prepared as follows: Two grams of the pyridinium bromide hydrobromide in ml. of water was added to a hot solution of 4.4 grams of sodium embonate in 75 ml. of water. The l-(2-ethyl-4- amino-S-pyridmidylmethyl)-2-methyl pyridinium embonate precipitated immediately, and was collected by filtration and washed with Water. It was recrystallized from 100 ml. of methanol to give substantially pure material, melting point 200 C. (dec.).

C. 3.01 grams of l(2-ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride was dissolved in 10 ml. of water. To this solution was added 3.5 grams of naphthalene-1,5-disulfonic acid. The solution remained clear for a few seconds and then crystallization of the naphthalene disultonate quaternary salt began. The reaction mixture was allowed to stand in ice water for two hours. At the end of which time, the 1- (2 ethyl-4-amino-S-pyrimidylmethyl)-2-methy1 pyridinium-l,5-naphthalene disulfonate was recovered by filtration and washed with water. It was obtained as colorless crystals, melting point 266-268 C. (dec.).

D. Fifteen grams of Amberlite IR-l20 resin was charged to a glass tube to give a resin column of about 18 cm. in height. The resin was converted to the hydrogen form by washing with dilute hydrochloric acid and then with water until the effluent had a pH of 5-6.

7.52 grams of 1-(2-ethyl-4-amino-S-pyrimidylmethyl)- Z-methyl pyridinium chloride hydrochloride in 325 ml. of water was passed over the column for a period of about two hours. The resin was then washed with 125 ml. of water and dried in vacuo at 50 C. By analysis of the efiluent it was determined that 7.15 grams of the pyridinium salt was absorbed on the resin. There was obtained 19.87 grams of 1-(2-ethyl-4-amino-5-pyrimidylmethyl)- Z-methyl pyridinium polystyrenesulfonate. N: calc"d, 6.0%; Found: 6.22%.

EXAMPLE 11 grams of 2amyl-4-amino-S-bromomethyl pyrimidine dihydrobromide in ml. of methanol was mixed with 15 ml. of oc-PlCOllHG. 100 ml. of isopropanol was added to the reaction mixture and the whole allowed to stand overnight at room temperature. Ether was then added to complete crystallization and the crystals of l-(2-amyl- 4-amino-S-pyrimidylmethyl)-2-methyl pyridinium bromide hydrobromide recovered by filtration. On recrystallization from ethanol-ether, the product melted at 205 C. (dec.).

EXAMPLE 12 To a suspension of 20 grams of 2-n-propyl-4-amino'- S-bromomethyl pyrimidine dihydrobromide in 120 ml. of acetonitrile was added 14 ml. of 4-methylpyridine. The mixture became clear after shaking for a few minutes and in a short time crystals began to form. The reaction mixture was allowed to stand overnight. The resulting crystals of 1-(2-n-propyl-4-amino-5-pyrimidylmethyl)-4- methyl pyridinium bromide hydrobromide were recovered by filtration and recrystallized from methanol-acetone, melting point 258260 C. (dec.).

When the above procedure was repeated using 16 ml. of 4-ethylpyridine, there was obtained 1-(2-n-propyl-4- amino-S-pyrimidylmethyl)-4-ethyl pyridinium bromide hydrobromide, melting point 258-260" C.; and when 18 ml. of 4-n-propylpyridine were employed, 1-(2-n-propyl- 1% 4-amino 5-pyrimidylmethyl)*4-n-propyl pyridinium bro= mide hydrobromide, melting point 244-246 C. (dec.), was obtained.

When the above procedure was carried out employing 20 grams of 2-ethyl-4-amino-5-bromethyl pyrimidine dihydrobromide and 14 ml. of B-methylpyridine as the reactants, there were recovered 1-(2-ethyl-4-amino-5-pyrimidylmethyl)-3-methyl pyridinium bromide hydrobromide, melting point 281-283 C. (dec.).

EMMPLE 13 To a stirred suspension of 20 grams of 2-ethyl-4-amino- S-pyrimidylmethyl bromide dihydrobromide in 125 ml. of acetonitrile was added 25 ml. of 4-methylpyridine.

The mixture became very warm and there was nearly a,

complete solution when a new solid began to crystallize. After standingfor 16 hours, 19 grams of l-(2-ethyl-4- amino-S-pyrimidylrhethyl)-4-rnethyl pyridinium bromide hydrobromide was obtained. 0n recrystallizing from methanol-acetone it had melting point 252 C. (dec.).

EXAMPLE 14 To a stirred suspension of 20 grams of 2-ethyl-4-amino- 5-pyrimidylmethyl bromide dihydrobromide, in ml. of acetonitrile was added 30 ml. of 4-n-propyl pyridine. When nearly all the solid has dissolved a new crystalline material began to crystallize. After 16 hours the crys talline product, 1-(2-ethyl-4-amino-S-pyrimidylmethyl) 4-propyl pyridinium bromide hydrobromide, was collected. It was recrystallized from ethanol-acetone, melting point 246 C. (dec.).

EXAMPLE 16 The 2-loweralkyl-4-amino-5-halomethyl pyrimidines employed in making the quaternary compounds of this invention are prepared by methods described in the literature or in the following manner:

A. 2-l0weralkyl-4-amino-5-cyanopyrimidina-A slurry of 54.7 grams of butyramidine hydrochloride and 55 ml. of absolute ethanol is treated at about 5 C. with a solution of 11 grams of sodium in 220 ml. of absolute ethanol. The resulting solution is added with stirring at iii-12 C. over a thirty minute period to 53.7 grams of ethoxymethylenemalononitrile in 54 ml. of absolute ethanol. The resulting mixture is stirred at 0 C. for six hours and then at room temperature for about 12 hours. The mixture is then filtered, evaporated to dryness in vacuo and the residue treated with water. The alcoholic and aqueous solution precipitates are combined, washed with Water and dried in air. The product is recrystallized from alcohol to give Z-n-propyl-4-amino-S-cyanopyrimidine, melting point 158-160 C.

When the above reaction is carried out with isobutyramidine there is obtained 2-isoprepyl-4-amino-5-cyanopyrimidine, melting point ISO-151 C. When amylamidine is employed as starting material the end product is 2-butyl-4-amino-S-Cyanopyrirnidine, melting point 143- 147 C. When hexylami'dine is utilized as starting compound, there is obtained 2-amyl-4-amino-S-cyanopyrimidine,-melting point 149-150" C.

B. 2-l0weralkyl-4-dmin0-5-amin0methy[pyrimidine dihydr0chl0ride.16.2 grams of 2-n-propyl-4-amino-5-cyanopyrimidine is reduced at about 40 lbs. pressure in 200 ml. of methanol in the presence of 26 grams of ammonia and one tablespoon of Raney nickel. The drop in pressure is about 36.5 lbs. The reaction mixture on completion of the reduction is concentrated to a syrup after filtering off the catalysts. The residue thus obtained is acidified with dilute hydrochloric acid and concentrated to a crystalline mass. On recrystallization from methanolacetone there is obtained 2-n-propyl-4-amino-5-aminomethylpyrimidine dihydrochloride, melting point 220-222 C.

When the 2-isopropyl, 2-butyl and 2-amyl-4-amino-5- cyanopyrimidines obtained as described above are used as starting materials in this reduction, there are obtained respectively 2-isopropyl-4-amino-5 aminomethylpyrirnidine dihydrochloride, melting point 257260 C., 2-butyl- 4-amino-5-aminomethylpyrimidine dihydrochloride, melting point 221223 C., and 2-amyl-4-amino-5-arninomethylpyrimidine dihydrochloride, melting point 188- 189 C.

C. 2-l0weralkyl-4-amin0-5-hydr0xy methyl pyrimidine. -Twelve grams of 2-n-propyl-4-amino-5-amino methylpyrimidine dihydrochloride in 50 ml. of water is treated at 50-60 C. with a solution of 3.5 grams of sodium nitrite in 30 ml. of water. The sodium nitrite is added dropwise over a 45 minute period. The heating is continued for an additional two hours, and the reaction mixture then made alkaline with sodium carbonate and extracted with butanol. The butanol extract is evaporated to dryness and the residue crystallized from acetone to give 2n-propyl-4-amiuo- S-hydroxy methylpyrimidine, melting point 115-6 C.

When the 2-isopropyl, Z-butyl and 2-amyl-4-amino-5- amino methylpyrimidine dihydrochlorides obtained as in part B above are utilized in this reaction in place of the 2-n-propy1 compound, there are obtained 2-isopropyl-4- amino-S-hydroxy methylpyrimidine, 2-butyl-4-amino-5- hydroxy methylpyrirnidine and 2-amyl-4-amino-5-hydroxy methylpyrimidine.

D. Z-loweralkyl-4-amina-5-br0mo m'ethylpyrimidz'ne. The 2-n-propyl-4-amino-5-hydroxy methylpyrirnidine obtained in part C above is dissolved in 15 ml. of acetic acid saturated with hydrogen bromide, and the mixture allowed to stand at room temperature for about 15 hours. 2-npropyl-4-amino-5-bromo methylpyrimidine dihydrobromide crystallizes and is recovered byfiltration and Washed with ether. The material is substantially pure and may be used directly in preparing the quaternary salts of this invention.

The other 2-loweralkyl-4-amino-S-hydroxy methylpyrimidines described above are treated in like manner with hydrogen bromide to give 2-ispropyl-4-amino-S-bromo methylpyrimidine dihydrobromide, melting point 191- 2 C., 2 butyl-4-amino-5-bromo methylpyrimidine dihydrobromide, melting point 145150 C., and 2-amyl-4- amino-S-bromo methylpyrimidine dihydrobromide.

EXAMPLE 17 When the quaternary salts of Examples 12 through 15 are treated with hydrochloric acid by the method of Example 6, the corresponding chloride hydrochloride quaternary salts are obtained.

Any departure from the above description which conforms to the present invention is intended to be included within the scope of the claims.

What is claimed is:

1. A composition that comprises a pyridinium quaternary salt of the formula orn-is X intimately dispersed in an inert edible carrier, wherein R is a lower alkyl group having at least two carbon atoms, R is lower alkyl, X is an anion, n has a value of 1 to 3 inclusive, and b and c are positive numbers having values such that the positive charge of b moles of cation is neutralized by 0 moles of anion X.

2. A poultry feed supplement composition that comprises an edible inert carrier having dispersed therein from about 1% to about 40% by weight of a pyridinium quaternary salt of the formula wherein R is a lower alkyl group having at least two carbon atoms, R is a lower alkyl group, and X is a nontoxic anion.

4. A composition useful against coccidiosis that comprises an edible carrier having intimately dispersed therein from about 1% to about 40% by weight of a 1-(2-lower alkyl 4 amino 5 pyrimidylmethyl) 2 methyl pyridinium quaternary salt, wherein the anion of the salt is a non-toxic anion and the lower alkyl radical contains at least two carbon atoms.

5. A composition useful against coccidiosis that comprises an inert edible carrier having intimately dispersed therein from about 1% to about 40% by weight of 1-(2- ethyl-4-amino-5-pyrimidylmethyl)-2-methyl pyridinium chloride hydrochloride.

6. A composition useful against coccidiosis that comprises an inert edible carrier having intimately dispersed therein from about 1% to about 40% by weight of 1-(2- propyl-4-arnino-5-pyrimidyhnethyl)-2-methyl pyridinium chloride hydrochloride.

7. A poultry feedstuff containing a 1-(2-loWer-alkyl- 4-amino-5-pyrimidylmethyl)-lower alkyl quaternary salt, wherein the lower alkyl group at the 2-position of the pyrimidine ring contains at least two carbon atoms, and the anion of the quaternary salt is a non-toxic anion.

8. An animal feedstuff having dispersed therein about 0.0005 to 0.05% by weight of pyridinium quaternary salt of the formula onriv wherein R' is a lower alkyl radical having at least two carbon atoms, R is a lower alkyl group, X is a non-toxic anion, n has a value of 1 to 3 inclusive, and b and c are positive numbers having values such that the positive charge of b moles of cation is neutralized by 0 moles of anion X. p

9. A water-soluble powder comprising an edible watersoluble carrier having dispersed therein from about 1.0% to about 25% by Weight of a quaternary salt of the formula 13 wherein R is a lower alkyl radical having at least two carbon atoms, R is a lower alkyl group, X is a non-toxic anion, n has a value of l to 3 inclusive, and b and c are positive numbers having values such that the positive charge of b moles of cation is neutralized by 0 moles of anion X.

10. An animal feed having dispersed therein from about 0.0005 to about 0.05% by weight of an acid addition salt of a 1-(2-loweralkyl-4-amino-5-pyrimidylmethyl)-2-loweralkyl pyridinium quaternary salt, wherein the lower alkyl group at the 2-position of the pyrimidine ring contains at least two carbon atoms.

11. An animal feedstuff having dispersed therein from about 0.0005% to about 0.05 by weight of 1-(2-ethyl-4- amino-S-pyrimidylmethyl) -2-methyl pyridiniurn halide hydrohalide.

12. An animal feedstufi having dispersed therein from about 0.0005 to about 0.05 by weight of 1-(2-propyl- 4-amino-5-pyrimidylmethyl)-2-methyl pyridinium halide hydrohalide.

13. An animal feedstuif having dispersed therein from about 0.0005% to about 0.05% by weight of l-(2-propyl- 4-amino-5-pyrimidylmethyl)-2 methyl pyridinium chloride hydrochloride.

14. An animal feed having dispersed therein from about 0.0005 to about 0.05% by weight of an acid addition salt of a 1-(2-loweralkyl-4-amino-S-pyrimidylmethyl)-4- loweralkyl pyridinium quaternary salt, wherein the lower alkyl group at the 2-position of the pyrimidine ring contains at least two carbon atoms.

15. A composition comprising an inert edible carrier having dispersed therein a l-(2-loweralkyl-4-arnino-5- pyrimidylmethyl)-loweralkyl pyridinium quaternary salt, wherein the lower alkyl group at the 2-position of the pyrimidine ring contains at least two carbon atoms, and the anion of the quaternary salt is a non-toxic anion.

References Cited in the file of this patent Fujita: J.B.C. 196, p. 297-303 and 313-320, 1952.

'UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,020,200 February 6, 1962 Edward F. Rogers et a1.

3 5 hereby certified that error appears in the above numbered patent requiring correction and that the said Letters Patent should read as corrected below- Column 12, line 47, before "quaternary" insert pyridinium Signed and sealed this 14th day of August 1962 (SEAL) Attest:

ERNEST W, SWIDER DAVID L. LADD Attesting Officer Commissioner of Patents 

2. A POULTRY FEED SUPPLEMENT COMPOSITION THAT COMPRISES AN EDIBLE INERT CARRIER HAVING DISPERSED THEREIN FROM ABOUT 2% TO ABOUT 40% BY WEIGHT OF A PYRIDINIUM QUATERNARY SALT OF THE FORMULA 